Pulsed field gel electrophoresis (PFGE) is regarded as the "third-generation molecular method" and the "gold standard" for bacterial typing. It is one of the practical methods that enable researchers to detect, identify and classify the genome of micro and macro organisms accurately and sensitively. Creative Biolabs is a leading provider of custom services for live biotherapeutic product (LBP) development. Our team applies decades to offer tailored, flexible solutions for microbial identification matched to your needs.
PFGE permits to resolve mixes of DNA molecules larger than 20 kb and up to 5 Mb in agarose gels. PFGE separates DNA based on the alternating of the two directions in which electric fields are applied, and the ability of the DNA molecule to reorient itself in the new direction in which the electric field is applied. The time consumed by the molecules in the reorientation depends on their sizes. Due to the advantages of PFGE in differentiating bacterial subtypes, PFGE is mainly used in bacterial DNA fingerprinting.
CHEF uses the positive electrode to the negative electrode at the angle of orientation 120 degrees. In this system, the size, position, coordination, stability, and continuity of electric fields are precisely controlled, which allows DNA fragments of different sizes to be separated.
Working with FIGE is easy and simple. FIGE is very popular nowadays because it can separate smaller parts. FIGE offers an acceptable resolution of over 800 Kb.
The method is used to detect DNA double-strand breaks (DSBs) by reverse asymmetric field gel electrophoresis.
RGE has a single electric field and can be separated directly because only one set of electrodes is used.
1. Preparation of Bacterial Cells
2. Preparation of Agarose Plugs
3. Plug Lysis
4. Plug Washing
5. Restriction Enzyme Digestion
6. Preparing and Running the Gel
7. Staining and Documentation
8. Data Analysis
Fig.1 A general scheme of the process of preparation of chromosome-sized immobilized DNA in agarose plugs, followed by PFGE analysis. (Lopez, 2019)
PFGE is used in the following areas:
PFGE is a powerful tool for subtyping bacteria. A good subtyping method should have five main characteristics: typability, reproducibility, discriminability, ease of implementation, and interpretability, and the PFGE method meets all these requirements. If you are developing LBP products and progressing to strain identification and strain typing, you can fully rely on Creative Biolabs’ PFGE service. Our expert scientists and talented technical support team have got you covered all the way. Please contact us for more information.
For Research Use Only. Not intended for use in food manufacturing or medical procedures (diagnostics or therapeutics). Do Not Use in Humans.